TRANSGENICS AND GENETIC CONSTRUCTS SHARED RESOURCE

转基因和基因构建共享资源(TGCSR)在目前的大部分资金中 Period提供三项主要服务：(1)通过DNA注射在近交系、混交系中产生转基因小鼠 近交系和近交系小鼠；(2)培养和转染ES细胞以产生“敲除”的ES克隆； 以及(3)利用转基因ES细胞培育“基因敲除”小鼠。平均而言，TGCSR 每年产生17个新的小鼠模型，由大约11个转基因小鼠品系组成 单个DNA构建和6个ES细胞来源的突变小鼠系。TGCSR提供了各种其他 服务，包括病原体感染转基因小鼠品系的重新衍生，冷冻保存 转基因小鼠胚胎和精子，帮助饲养和转基因小鼠的基因分型 并协助进行特殊手术或为实验目的进行静脉注射。最近， TGCSR已经进行了重组，以最大限度地提高效率，并扩大对达特茅斯调查人员的服务。 转基因和遗传构建共享资源(TGCSR)提供额外的主要服务 在大肠杆菌和酵母中使用“重组工程”的基因构建，提纯DNA用于对小鼠或胚胎进行基因分型 细胞，对小鼠或ES细胞进行PCR或Southern blotting，并对ES克隆进行核型分析。这个 TGCSR服务从计划一种新的老鼠模型开始，以转基因动物结束， 调查员没有进一步的劳动。这些扩展的、从开始到结束的服务将支持这一代 达特茅斯大学的研究人员在他们的实验室里不是分子生物学家的新的实验模型。在……里面 以便更有效地利用资源并响应NCI的当前指令，该指令要求 癌症中心寻求区域合作，并在可行的情况下联合利用共享资源， 莫里斯棉花癌症中心已与杰克逊实验室(JAX)达成协议，将拥有 由JAX的转基因共享资源完成对TGCSR的小鼠胚胎操作。都是以前的事 提供的不涉及操纵小鼠胚胎的服务将继续在 TGCSR的NCCC。在这项新安排下，TGCSR总监将成为所有人的渠道 达特茅斯调查人员与JAX之间的通信，为JAX提供试剂，计费 调查人员，以及对日本宇宙航空研究开发机构的经济补偿。日本宇宙航空研究开发机构在以下领域的丰富经验和资源 小鼠遗传学将为NCCC的研究人员提供更好的服务，而且由于JAX直接做出了 每周将无病原体的小鼠送到达特茅斯，转基因小鼠的运动将是有效的 对所有人来说都很方便。通过与世界上最先进的老鼠遗传学研究机构JAX合作，以及通过 将我们的服务扩展到包括基因构建生产、核型分析和基因分型，我们将更好地 为NCCC调查人员服务。

The Transgenics and Genetic Constructs Shared Resource (TGCSR) during much of the current funding period provided three main services: (1) generation of transgenic mice by DNA injection in outbred, mixed inbred and inbred mouse strains; (2) culture and transfection of ES cells to generate "knockout" ES clones; and (3) production of "knockout" mice using genetically modified ES cells. On average, the TGCSR generated 17 novel mouse models per year, consisting of approximately 11 transgenic mouse lines from individual DNA constructs and 6 ES cell-derived mutant mouse lines. The TGCSR provided various other services, including rederivation of pathogen-infected transgenic mouse lines, cryopreservation of transgenic mouse embryos and sperm, assistance with husbandry and genotyping of transgenic mouse lines, and assistance with special surgeries or IV injections for experimental purposes. Recently, the TGCSR has been reorganized to maximize efficiency and expand services to Dartmouth investigators. Transgenic and Genetic Construct Shared Resource (TGCSR) provides the additional major services of genetic constructs using "recombineering" in E. coli and yeast, purifying DNA for genotyping mice or ES cells, performing PCR or Southern blotting to genotype mice or ES cells, and karyotyping ES clones. The TGCSR services start with planning a new mouse model and end with genetically modified animals, with no further labor from the investigator. These expanded, start-to-finish services will support the generation of new experimental models by Dartmouth investigators who are not molecular biologists in their labs. In order to more efficiently utilize resources and respond to current directives from the NCI requesting that cancer centers seek regional collaboration and combine utilization of shared resources whenever feasible, Morris Cotton Cancer Center has entered into an agreement with the Jackson Laboratory (JAX) to have the mouse embryo manipulation for the TGCSR done by the transgenic shared resource at JAX. All previously provided services that do not involve manipulation of mouse embryos will continue to be performed at NCCC by the TGCSR. Under this new arrangement, the TGCSR director will be the conduit for all communications between Dartmouth investigators and JAX, for providing reagents to JAX, billing investigators, and financially compensating JAX. The vast experience and resources of JAX in the area of mouse genetics will provide even better service for NCCC investigators, and, since JAX makes a direct delivery of pathogen-free mice to Dartmouth every week, the movement of transgenic mice will be efficient and convenient for all. By partnering with JAX, the world's foremost facility for mouse genetics, and by expanding our services to include genetic construct production, karyotyping, and genotyping, we will better serve NCCC investigators.